Isolation of Human Small Extracellular Vesicles and Tracking of their Uptake by Retinal Pigment Epithelial Cells In Vitro.

Marcu, Irene C.; Eberhard, Naja; Yerly, Anaïs; Balmer, Verena; Hemphill, Andrew; Mogel, Helga; Gaschen, Véronique; Stoffel, Michael H.; Bluteau, Jasmin (2020). Isolation of Human Small Extracellular Vesicles and Tracking of their Uptake by Retinal Pigment Epithelial Cells In Vitro. International journal of molecular sciences, 21(11), pp. 1-19. MDPI 10.3390/ijms21113799

[img]
Preview
Text
ijms-21-03799(1).pdf - Published Version
Available under License Creative Commons: Attribution (CC-BY).

Download (3MB) | Preview

Small extracellular vesicles (EVs) are among the most frequently investigated EVs and play major roles in intercellular communication by delivering various cargo molecules to target cells. They could potentially represent an alternative delivery strategy to treat ocular toxoplasmosis, a parasitosis affecting the retinal pigment epithelium (RPE). To date, the uptake of human small EVs by RPE cells has never been reported. In this study, we report on the intracellular uptake of fluorescently labelled human urine and fibroblast-derived small EVs by human RPE cells. In summary, both dye-labelled urinary small EVs and small EVs obtained from fibroblasts stably expressing membrane-bound green fluorescent protein were successfully internalized by RPE cells as revealed by immunohistochemistry. In recipient ARPE19 cells, BODIPY-labelled small EVs were found in close vicinity to the parasite Toxoplasma gondii. Additionally, an ultrastructural method was enabled to distinguish between labelled exogenous and endogenous small EVs within target cells.

Item Type:

Journal Article (Original Article)

Division/Institute:

05 Veterinary Medicine > Department of Infectious Diseases and Pathobiology (DIP) > Institute of Parasitology
05 Veterinary Medicine > Department of Clinical Research and Veterinary Public Health (DCR-VPH) > Veterinary Anatomy
09 Interdisciplinary Units > Microscopy Imaging Center (MIC)
05 Veterinary Medicine > Department of Clinical Research and Veterinary Public Health (DCR-VPH)

UniBE Contributor:

Marcu, Irene Cristina; Eberhard, Naja; Yerly, Anaïs Elodie; Balmer, Vreni; Hemphill, Andrew; Mogel-Allemann, Helga Maria; Gaschen, Véronique; Stoffel, Michael Hubert and Bluteau, Jasmin Maria

Subjects:

600 Technology > 630 Agriculture
500 Science > 570 Life sciences; biology
600 Technology > 610 Medicine & health

ISSN:

1422-0067

Publisher:

MDPI

Language:

English

Submitter:

Vanessa Alice Blum

Date Deposited:

09 Jun 2020 09:02

Last Modified:

09 Jun 2020 09:02

Publisher DOI:

10.3390/ijms21113799

PubMed ID:

32471212

Uncontrolled Keywords:

small extracellular vesicles; BODIPY; PKH; EV; retinal pigment epithelium; T. gondii

BORIS DOI:

10.7892/boris.144513

URI:

https://boris.unibe.ch/id/eprint/144513

Actions (login required)

Edit item Edit item
Provide Feedback